The Protein Purification Core is a shared facility to provide requisite amounts of purified, biologically active MDA-7 protein for utilization as a key reagent in the hypothesis-driven studies proposed in the various Projects of this proposal. Utilization of protein for therapy of prostate cancer (Project 1, Paul B. Fisher), malignant glioma (Project 2, Paul Dent) and ovarian cancer (Project 3, David T. Curiel) will be studied based on previous data showing that a bacterially expressed GST-fusion of MDA-7 (GST-MDA-7) shows the same cancer cell specific apoptosis inducing activity observed after infection with Ad.mda-7. The core will initially provide required preparative scale (milligram quantities) of purified, biologically tested protein for each of these projects in addition to control protein including unfused GST. In addition, the core will establish by constructing vectors, standardizing and scaling up a procedure based on an insect cell baculovirus expression system, for production of preparative scale MDA-7 protein with a minimal fusion tag. This protein will have specific utility in tumor infusion studies based on animal models, proposed in Projects 1, 2 and 3. The core will also be in a position to generate additional expression vector systems and produce purified protein, to meet specific reagent needs to the component Projects of the PPG. These could include specific purified components of the Ras and MAP kinase pathways used in in vitro tests for activation of specific types of a signal transduction cascades that might be triggered by mda-7. The centralization of these activities as a shared resource will enable these reagents to be produced in an efficient and cost-effective manner by capitalizing on existing expertise in this area.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Program Projects (P01)
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Subcommittee G - Education (NCI)
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Virginia Commonwealth University
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Menezes, Mitchell E; Bhoopathi, Praveen; Pradhan, Anjan K et al. (2018) Role of MDA-7/IL-24 a Multifunction Protein in Human Diseases. Adv Cancer Res 138:143-182
Pradhan, Anjan K; Emdad, Luni; Das, Swadesh K et al. (2017) The Enigma of miRNA Regulation in Cancer. Adv Cancer Res 135:25-52
Shapiro, Brian A; Vu, Ngoc T; Shultz, Michael D et al. (2016) Melanoma Differentiation-associated Gene 7/IL-24 Exerts Cytotoxic Effects by Altering the Alternative Splicing of Bcl-x Pre-mRNA via the SRC/PKC? Signaling Axis. J Biol Chem 291:21669-21681
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Talukdar, Sarmistha; Emdad, Luni; Das, Swadesh K et al. (2015) Noninvasive approaches for detecting and monitoring bladder cancer. Expert Rev Anticancer Ther 15:283-94
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Sarkar, Siddik; Quinn, Bridget A; Shen, Xuening et al. (2015) Reversing translational suppression and induction of toxicity in pancreatic cancer cells using a chemoprevention gene therapy approach. Mol Pharmacol 87:286-95
Das, Swadesh K; Menezes, Mitchell E; Bhatia, Shilpa et al. (2015) Gene Therapies for Cancer: Strategies, Challenges and Successes. J Cell Physiol 230:259-71
Azab, Belal M; Dash, Rupesh; Das, Swadesh K et al. (2014) Enhanced prostate cancer gene transfer and therapy using a novel serotype chimera cancer terminator virus (Ad.5/3-CTV). J Cell Physiol 229:34-43
Menezes, Mitchell E; Das, Swadesh K; Emdad, Luni et al. (2014) Genetically engineered mice as experimental tools to dissect the critical events in breast cancer. Adv Cancer Res 121:331-382

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